Dengue virus HA antigens in infected cells from Aedes albopictus clone C6/36 can be produced easily. Their titers that prepared were from 64 to 128 times higher than from baby mouse brain. The method is recommended as an initial step in the concentration of Dengue virus propagated in tissue cullure, as it is simple, rapid and inexpensive
Dengue Virus ; Hemagglutinins

PURPOSE: Nitrocellulose membrane–based filtration system (NCFS) is widely used for protein concentration. In this study, we applied NCFS for production of virus-like particle (VLP) as a vaccine candidate and evaluated yield property and immunogenicity. MATERIALS AND METHODS: Influenza VLPs were generated by baculovirus-insect cell protein expression system. NCFS and sucrose gradient ultracentrifugation were used for purification of VLP. Immunogenicity of VLP was evaluated by animal experiment. RESULTS: Influenza VLPs expressing hemagglutinin (HA) and neuraminidase proteins derived from highly pathogenic influenza virus (H5N8) were effectively produced and purified by NCFS. HA activity of VLP which correlated with antigenicity was well conserved during multiple purification steps. This NCFS based purified VLPs induced influenza virus–specific antibody responses. CONCLUSION: Our results indicate that the influenza VLP vaccine could be prepared by NCFS without loss of immunogenicity and elicit antigen-specific immune responses.
Animal Experimentation ; Antibody Formation ; Baculoviridae ; Collodion* ; Filtration* ; Hemagglutinins ; Influenza, Human* ; Membranes* ; Neuraminidase ; Orthomyxoviridae ; Sucrose ; Ultracentrifugation ; Vaccines

We report a case of cold hemagglutinin disease associated with Mycoplasma pneumoniae infection treated with cord blood transfusion. Cold hemagglutinin disease is a hemolytic anemia most commonly associated with cold-reactive autoantibody with anti-I specificity. On the basis of the fact that the level of I antigen on cord red blood cells is extremely low, a six year old male patient was transfused with 60 mL of ABO blood type-matched, cord blood. No complication from the transfusion was observed. Due to the deficiency in cord blood supply, filtered irradiated RBC 100 mL was transfused three times thereafter. The hemoglobin level began to increase from the fifth hospital day. The patient was discharged without additional transfusion on the eleventh hospital day. No remarkable complications were noted at the time of discharge.
Anemia, Hemolytic ; Erythrocytes ; Fetal Blood* ; Hemagglutinins* ; Humans ; Male ; Mycoplasma pneumoniae* ; Pneumonia, Mycoplasma* ; Sensitivity and Specificity

In late March of 2009, an outbreak of influenza in Mexico, was eventually identified as H1N1 influenza A. In June 2009, the World Health Organization raised a pandemic alert to the highest level. More than 214 countries have reported confirmed cases of pandemic H1N1 influenza A. In Korea, the first case of pandemic influenza A/H1N1 infection was reported on May 2, 2009. Between May 2009 and August 2010, 750,000 cases of pandemic influenza A/H1N1 were confirmed by laboratory test. The H1N1-related death toll was estimated to reach 252 individuals. Almost one billion cases of influenza occurs globally every year, resulting in 300,000 to 500,000 deaths. Influenza vaccination induces virus-neutralizing antibodies, mainly against hemagglutinin, which provide protection from invading virus. New quadrivalent inactivated influenza vaccine generates similar immune responses against the three influenza strains contained in two types of trivalent vaccines and superior responses against the additional B strain.
Antibodies ; Hemagglutinins ; Influenza Vaccines ; Influenza, Human* ; Korea* ; Mexico ; Pandemics* ; Vaccination ; Vaccines ; World Health Organization

The in vivo and in vitro humoral and cell-mediated immune responses of lymphocytes of BALB/c mouse exposed to mercury chloride(HgC12) were investigated. In vitro exposure of the splenocytes to mercury chloride produced overt cytotoxicity in 3 hours period. The IC50(the concentration required to inhibit a splenocyte viability by 50%) for mercury chloride was >0.1mM for cytotoxicity. In vivo mercury chloride exposed mice were significantly depressed delayed type hypersensitivity(DTH) response to sheep red blood cells(SRBC) in a dose-dependent manner compared with control group. Mercury chloride inhibited the proliferative responses of splenocytes to lipopolysaccharide. pokeweed mitogen, concanavalin A and phytohemagglutinin in a dose-dependent manner. Hemagglutinin response to SRBC in mercury chloride exposed mice was significantly depressed in a dose-dependent manner compared with control group. After 7 weeks of mercury chloride exposure in vivo. mercury chloride induced an increase of nonspecific serum IgG1 and IgE levels in BALB/c mice.
Animals ; Concanavalin A ; Hemagglutinins ; Immunoglobulin E ; Immunoglobulin G ; Lymphocytes ; Mice* ; Phytolacca americana ; Sheep

OBJECTIVETo construct and identify the Porphyromonas gingivalis(Pg)ATCC33277 hemagglutinin-2(HA-2)-deficient mutant.

METHODSThe genomic DNA of Pg was isolated from PgATCC33277. The up/down stream genes of HA-2-HA(u), HA(l) were amplified by PCR, and inserted into pSY118 separately which contains a 2.1 kb antibiotic resistance ermF-ermAM cassette. The resultant recombinant plasmid-pSY118-HA was linearized as the gene targating fragment HA-ermF-ermAM and used in the electroporation of PgATCC33277. The Pg HA-2-deficient mutant was screened by allelic exchange. The test of aggregation of red blood cells was used to investigate the function change between PgHA2-deficient mutant and the wild type of PgATCC33277.

RESULTSThe PgHA-2-deficient mutant was identified by PCR. The ability of Pg HA-2-deficient mutant to aggregate red blood cell was significantly decreased compared with the wild type.

CONCLUSIONSHA-2-deficient mutant of Pg ATCC33277 was constructed successfully, which lays a foundation for further study of its biological function.

Alleles ; Erythrocyte Aggregation ; Hemagglutinins ; genetics ; Polymerase Chain Reaction ; Porphyromonas gingivalis ; genetics

OBJECTIVETo determine the sequence of the active peptide derived from recombinant hemagglutinin (rHA-2) of Porphyromonas gingivalis (Pg).

METHODSThe HA-2 gene from PgATCC33277 was cloned, expressed in Escherichia coli (Ec) BL21 (DE3), and purified. The purified recombinant protein was evaluated for its ability to bind hemin-linked agarose. The active peptide was subjected to endoproteinase-mediated sequence analysis.

RESULTSThe protein expressed in Ec BL21 (DE3) was identified as PgHA-2 by plasmid sequence analysis, Western blotting, and mass spectrometry. The recombinant protein was confirmed functional by its ability to bind hemin. The sequence of the active peptide of rHA-2 was determined to be DHYAVMISKTGTNAG.

CONCLUSIONSThe availability of sequence of the active peptide of rHA-2 provides a foundation for the development of immunoprophylactic and therapeutic agents against this human pathogen.

Bacterial Proteins ; chemistry ; genetics ; Hemagglutinins ; chemistry ; genetics ; Porphyromonas gingivalis ; genetics ; Sequence Analysis, Protein

BACKGROUND: Community-acquired pneumonia(CAP) remains a leading cause of morbidity and mortality worldwide. Recently, the evolution of drug-resistant microorganisms has become a serious problem in CAP management. Specific antimicrobial therapy is the cornerstone of CAP management. However, obtaining an accurate etiologic diagnosis clinically is not easy and empirical antimicrobial treatment is usually administered prior to the correct microbiologic diagnosis. In this study, the clinical usefulness of empirical CAP treatment was investigated. METHODS: A total 35 cases were studied prospectively over a 16-month period in Mokpo Catholic Hospital from Dec. 1995 to Mar. 1997. The microbiologic diagnosis was made by sputum, blood culture, a specific serum antibody test and an immunologic study. RESULTS: The causative organisms were isolated in 10 (30%) out of 33 cases: 8 cases and 1 case on the sputum culture and blood culture respectively, and 1 case by an indirect hemagglutinin test. 12 cases had underlying diseases: pulmonary tuberculosis 4, alcoholism 4, diabetes mellitus 3, and liver cirrhosis 1. Antimicrobial treatment was given empirically and all cases recovered. CONCLUSION: A definite microbiologic diagnosis before commencing the appropriate treatment in CAP is not straightforward. Empirical therapy according to a clinical assessment is important and helpful. However, every effort to make the correct etiologic diagnosis should be taken.
Alcoholism ; Diabetes Mellitus ; Diagnosis ; Hemagglutinins ; Jeollanam-do* ; Liver Cirrhosis ; Mortality ; Pneumonia* ; Prospective Studies ; Sputum ; Tuberculosis, Pulmonary

OBJECTIVETo investigate the variations on hemagglutinin (H) gene of measles virus (MV) in Zhejiang province, and to analyze the differences with strains circulated both at home and abroad.

METHODSIn total, 33 MV strains isolated in Zhejiang province between 1999 and 2011 were collected.RNA of the isolated MV strains was extracted and the complete sequences on H gene were amplified using RT-PCR assay. The products were compared with the Chinese vaccine strain Shanghai-191, which was downloaded from GenBank, and other 95 different MV strains from all over the world.

RESULTS33 MV strains, isolated from the throat swab specimens collected from MV patients in Zhejiang province during 1999 to 2001, were used to conduct phylogenetic analysis with MV strains circulated in other areas of China during 1993 to 2007. The phylogenetic tree based on H gene sequences showed that all the Zhejiang MV strains located in H1a cluster, and no apparent time series and geographic restrictions were observed. Compared to the referenced vaccine strain Shanghai-191, the average variation rate on nucleotides and amino acids, and the evolutionary rate of H1a viruses from China during 2003 to 2011 were separately 5.15%, 4.44% and 5.81%, which were higher than the rates of H1a viruses during 1965 to 1993 (4.75%, 3.86% and 5.30%), and the rates of viruses during 1994 to 2002 (4.80%, 4.08% and 5.37%).However, the dn/ds ratios of strains within the three time periods were 0.19,0.21 and 0.23 respectively, which indicated that no evidence of positive selection was found on H1a MV strains during 1993 to 2011. A 24 stable amino acid variation sites on H gene was found between H1a viruses during 2003 to 2011 and the vaccine strain Shanghai-191. The largest variation occurred between vaccine and H1a strains, with 0.053 of the p-distance and 26-28 of amino acid mutations.However, only 15 stable amino acid variations were found between vaccine strain and genotype B3 or D4 strains.In addition, significant differences were found between H1a viruses and genotype B or D viruses, with 0.074 and 0.071 of p-distance and 27-33 of amino acid differences.

CONCLUSIONSignificant differences were found on H gene between MV strains subtype H1a and vaccine strains and other genotype strains. The variations were enlarged with the time coursing; therefore, the surveillance on variation of Chinese MV strains should be taken into account.

China ; epidemiology ; Genotype ; Hemagglutinins, Viral ; genetics ; Humans ; Measles ; epidemiology ; virology ; Measles virus ; classification ; genetics ; isolation & purification

PURPOSE: We conducted the study to evaluate the immunogenicity and safety of three component DTaP vaccine (Infanrix(R)) in a group of Korean healthy infants on a three-dose primary vaccination. And we compared the immunogenicity of this DTaP vaccine with two component DTaP vaccine which has been widely used in Korea. METHODS: We enrolled one hundred fifty one healthy infants aged 8-9 weeks. These infants were vaccinated at age 2, 4 and 6 months of age with three component DTaP vaccine. Solicited adverse events were actively monitored for 72 hours following each vaccination, and all adverse events after each vaccination were observed for three weeks. Anti-diphtheria toxoid Ab., anti-tetanus toxoid Ab., anti-pertussis toxin Ab., anti-filamentous hemagglutinin Ab., and anti-pertactin Ab. were measured using ELISA for assessing immunogenicity of study vaccine in 60 infants. Immunogenicity analysis of two component DTaP vaccine was performed with same methods in 14 infants as control. RESULTS: The seroconversion rates of anti-diphtheria toxoid Ab, anti-tetanus toxoid Ab. anti- filamentous hemagglutinin Ab. were 100% in both group. Seroconversion rate of anti-pertactin Ab in study group was 100%, but the rate in control group was 50%. However, geometric mean concentration of anti-pertussis toxin Ab. was higher in control group. Mild local and systemic reactions were observed within three days after vaccination, and no serious adverse events related study vaccine were happened during study period. CONCLUSIONS: Our study results suggest that three component DTaP vaccine (Infanrix(R)) is a well- tolerable and high immunogenic vaccine, especially anti-Pertactin Ab. of the study vaccine is very immunogenic. It can be available as routine DTaP vaccination in our infants.
Diphtheria-Tetanus-acellular Pertussis Vaccines* ; Enzyme-Linked Immunosorbent Assay ; Hemagglutinins ; Humans ; Infant* ; Korea ; Pertussis Toxin ; Vaccination